2026-10-06T09:17:58Z https://api.figshare.com/v2/oai

oai:figshare.com:article/34074603 2026-10-05T18:20:28Z category_1 category_4 category_7 category_12 category_14 category_15 category_46 category_122 category_132 category_134 portal_5 item_type_3 month_year_10_2026

<p>Primers used for qPCR.</p> Xuan Chen (158617) Chuang Li (2010226) Dongyue Xing (25317416) Jingjing Song (711775) Yun Liu (84346) Yuxin Liu (505575) Qin Huang (49441) Yu Kuang (763791) Xian Li (201841) Jinlei Zhang (3521606) Yuqi Zhang (286958) Nianzhi Zhang (8262480) Rulan Bai (120694) Changyong Cheng (1624090) Jin Yuan (120922) Jun Tang (32983) Rui Zhang (13940) Biophysics Biochemistry Medicine Cell Biology Molecular Biology Neuroscience Immunology Mental Health Infectious Diseases Virology milder histopathological lesions lysine residues k242 neuronal n2a cells epithelial pk15 cells subsequent productive replication facilitates neuronal replication facilitates efficient replication reducing nuclear accumulation div >< p requires efficient reactivation reduced viral loads tk ), encoded mouse infection model establishes lifelong latency affect tk stability reactivation </ p identify tk sumoylation viral replication identify tk reactivation model dividing cells vitro </ prv ), nuclear localization viral infection viral reactivation prv tk prv infection latent infection deficient tk translational regulation thymidine kinase subcellular localization small ubiquitin pseudorabies virus potential target markedly alters like modifier kinase activity k267 serving functional significance findings demonstrate biological function <div><p>Pseudorabies virus (PRV), an alphaherpesvirus, establishes lifelong latency in neurons and requires efficient reactivation for pathogenesis. Thymidine kinase (TK), encoded by UL23, is a key virulence factor essential for viral replication in non-dividing cells and for reactivation from latency; however, the regulatory mechanisms governing TK function remain poorly defined. Here, we identify TK as a substrate for small ubiquitin-like modifier (SUMO) modification and investigate its functional significance during PRV infection. We show that TK undergoes endogenous SUMOylation during viral infection is conjugated by SUMO-1, SUMO-2, and SUMO-3, with lysine residues K242 and K267 serving as the principal SUMO acceptor sites. Disruption of these SUMOylation sites does not affect TK stability and kinase activity but markedly alters its subcellular localization by reducing nuclear accumulation. Functionally, SUMOylation of TK is dispensable for viral replication in epithelial PK15 cells but facilitates efficient replication in neuronal N2a cells. Using an <i>in vitro</i> latency/reactivation model, we further show that SUMOylation-deficient TK does not affect the establishment of latent infection but markedly impairs viral reactivation and subsequent productive replication. In a mouse infection model, disruption of TK SUMOylation attenuates PRV pathogenicity, accompanied by reduced viral loads in the brain and milder histopathological lesions. Collectively, our findings demonstrate that SUMOylation regulates nuclear localization and biological function of PRV TK, thereby facilitating efficient neuronal replication, viral reactivation, and pathogenesis. These findings provide new insights into the post-translational regulation of alphaherpesvirus TK and identify TK SUMOylation as a potential target for controlling PRV infection.</p></div> 2026-10-05T18:20:02Z Dataset Dataset 10.1371/journal.ppat.1014653.t003 https://figshare.com/articles/dataset/_p_Primers_used_for_qPCR_p_/34074603 CC BY 4.0